Identification and characterization of 31 isolates of Borrelia burgdorferi (Spirochaetales, Spirochaetaceae) obtained from various hosts and vectors using PCR-RFLP and SDS-PAGE analysis

Abstract

Borrelia burgdorferi sensu lato, the etiologic agent of Lyme borreliosis, circulates between ticks and vertebrate hosts. Two main genospecies typically occur in the Czech Republic Borrelia garinii and Borrelia afzelii, transmitted generally by Ixodes ricinus (L., 1758) ticks. The aim of our study was to identify spirochaete isolates focusing on Borrelia burgdorferi acquired from different sources: vectors (ticks), potential vectors (mosquitoes, small mites) and hosts (wild rodents). In the years 1996-2001 a total of 2398 ticks, 72 mites (from wild rodents), 2700 mosquito adults, 1798 mosquito larvae and organ parts (kidney and spleen) of 216 wild rodents were collected from seven localities in the Czech Republic. A total of 31 spirochaete strains were isolated: 13 strains from ticks, 1 strain from mite (Haemogamasus sp.), 15 strains from rodents, 1 strain from mosquito adults and 1 strain from mosquito larva. For the genospecies identification of these isolates PCR, PCR-RFLP was used and their characterization was also performed by SDS-PAGE. By nested PCR method all except one isolated strains were detected as Borrelia burgdorferi s.l. Following PCR-RFLP molecular analysis results, tick isolates were identified as B. garinii and B. afzelii, the strain isolated from the mite was identified as B. afzelii. This is the first isolated strain of B.b.s.l. from a different mite of infraorder Parasitiformes than tick. All of rodent isolates were identified as B. afzelii; mosquito adult isolate was identified as B. afzelii. Larval isolate from mosquito is spirochaete, but does not belong to Borrelia burgdorferi sensu lato group.

Identification of Borrelia burgdorferi genospecies isolated from Ixodes ricinus ticks in the South Moravian region of the Czech Republic

“…In the presented results, the higher occurence of B. afzelii (70%) was expected according to previous studies from the Czech Republic. B. afzelii with B. garinii (10%) are the most abundant genospecies in the Czech Republic [25]. In the current results, genospecies B. valaisiana (8.6%), B. spielmanii (7.1%) and B. burgdorferi s. s. (4.3%) were also identified.…”

Is there a Lyme-like disease in Australia? Summary of the findings to date

“…In the northern hemisphere, B. burgdorferi s. l. has also been detected in Ixodidae (hard tick) of genera other than Ixodes [1] , [2] , [3] , [4] and even in haematophagous arthropods including lice [2] , fleas [4] , [5] , keds [2] , [6] , mites [7] , [8] , [9] , flies [10] , [11] , [12] and mosquitoes [9] , [13] , [14] . While the transmission capability of these arthropods remains undetermined, it does raise the possibility of Lyme transmission by arthropods other than Ixodes.…”

“…Typically, the disease first presents with an erythema migrans rash at the site of the tick bite, followed by flu-like symptoms and later by debilitating arthritic, dermatological and neurological manifestations. The bacteria are transmitted by Ixodes species ticks, although other Ixodidae ticks [1] , [2] , [3] , [4] and haematophagous arthropods [2] , [4] , [5] , [6] , [7] , [8] , [9] , [10] , [11] , [12] , [13] , [14] have been implicated in carrying the bacteria. Bacterial reservoirs of the disease are usually small mammals, birds and occasionally reptiles [15] , [16] .…”

Activity of the tickIxodes ricinusmonitored in a suburban park in Brno, Czech Republic, in association with the evaluation of selected repellents

“…positive samples from I. ricinus in this locality (Žákovská et al ). From these, a total of ten isolated strains (6.2%) were obtained, seven of which were identified as B. afzelii and three as B. garinii , using PCR‐RFLP (Žákovská et al ). The presence of B. garinii suggests a role for rodents in the circulation of B. burgdorferi s.l.…”

“…The DFM method revealed 50 samples positive for the presence of spirochaetes (8.4%). The DFM method is commonly used for the detection of spirochetes in ticks (Žákovská et al , Aleksejev et al , Stünzner et al , Hubálek et al ). A reason for finding fewer positive samples in PCR could be that our PCR sensitivity cannot detect a very low number of borreliae (from 0–10 per tick).…”